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甲醇固定饲养层改变了牛多能干细胞的多能性和代谢。

Methanol fixed feeder layers altered the pluripotency and metabolism of bovine pluripotent stem cells.

机构信息

State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia University, Hohhot, People's Republic of China.

Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Baotou, Inner Mongolia, People's Republic of China.

出版信息

Sci Rep. 2022 Jun 2;12(1):9177. doi: 10.1038/s41598-022-13249-3.

DOI:10.1038/s41598-022-13249-3
PMID:35654935
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9163156/
Abstract

The pluripotency maintenance of pluripotent stem cells (PSCs) requires the suitable microenvironment, which commonly provided by feeder layers. However, the preparation of feeder layers is time consuming and labor exhaustive, and the feeder cells treated with mitomycin C or γ-ray irradiation bring heterologous contamination. In this study, mouse embryonic fibroblasts (MEFs) were treated by methanol to generate chemical fixed feeder cells, and bovine embryonic stem cells F7 (bESC-F7) cultured on this feeder layer. Then the pluripotency and metabolism of bESC-F7 cultured on methanol-fixed MEFs (MT-MEFs) named MT-F7 was compared with mitomycin C treated MEFs (MC-MEFs). The results showed that bESC-F7 formed alkaline phosphatase positive colonies on MT-MEFs, the relative expression of pluripotent markers of these cells was different from the bESCs cultured on the MC-MEFs (MC-F7). The long-term cultured MT-F7 formed embryoid bodies, showed the ability to differentiate into three germ layers similar to MC-F7. The analyses of RNA-seq data showed that MT-MEFs lead bESCs to novel steady expression patterns of genes regulating pluripotency and metabolism. Furthermore, the bovine expanded pluripotent stem cells (bEPSCs) cultured on MT-MEFs formed classical colonies, maintained pluripotency, and elevated metabolism. In conclusion, MT-MEFs were efficient feeder layer that maintain the distinctive pluripotency and metabolism of PSCs.

摘要

多能干细胞（PSCs）的多能性维持需要合适的微环境，通常由饲养层提供。然而，饲养层的制备既费时又费力，用丝裂霉素 C 或 γ 射线处理的饲养细胞会带来异源污染。在这项研究中，用甲醇处理小鼠胚胎成纤维细胞（MEFs）以产生化学固定饲养细胞，并在这种饲养层上培养牛胚胎干细胞 F7（bESC-F7）。然后，将牛胚胎干细胞 F7 在甲醇固定 MEFs（MT-MEFs）上培养的多能性和代谢与用丝裂霉素 C 处理的 MEFs（MC-MEFs）培养的进行比较。结果表明，bESC-F7 在 MT-MEFs 上形成碱性磷酸酶阳性集落，这些细胞的多能性标记的相对表达与在 MC-MEFs 上培养的 bESCs 不同（MC-F7）。长期培养的 MT-F7 形成胚状体，表现出类似于 MC-F7 的向三个胚层分化的能力。RNA-seq 数据分析表明，MT-MEFs 使 bESCs 形成了调节多能性和代谢的基因的新型稳定表达模式。此外，在 MT-MEFs 上培养的牛扩增多能干细胞（bEPSCs）形成了典型的集落，保持了多能性，并提高了代谢水平。总之，MT-MEFs 是一种有效的饲养层，能维持 PSCs 的独特多能性和代谢。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/4d76bed3ab6e/41598_2022_13249_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/6f7d4e709f96/41598_2022_13249_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/57cb488f551d/41598_2022_13249_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/a1a60423ca40/41598_2022_13249_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/c7f8502b1ab8/41598_2022_13249_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/aaf16eb50b3e/41598_2022_13249_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/8523a360fadb/41598_2022_13249_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/6fd80257ff36/41598_2022_13249_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/4d76bed3ab6e/41598_2022_13249_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/6f7d4e709f96/41598_2022_13249_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/57cb488f551d/41598_2022_13249_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/a1a60423ca40/41598_2022_13249_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/c7f8502b1ab8/41598_2022_13249_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/aaf16eb50b3e/41598_2022_13249_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/8523a360fadb/41598_2022_13249_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/6fd80257ff36/41598_2022_13249_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29e8/9163156/4d76bed3ab6e/41598_2022_13249_Fig8_HTML.jpg

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